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地球上准微重力环境下高质量蛋白质晶体的原位实时生长观测。

In-situ and real-time growth observation of high-quality protein crystals under quasi-microgravity on earth.

作者信息

Nakamura Akira, Ohtsuka Jun, Kashiwagi Tatsuki, Numoto Nobutaka, Hirota Noriyuki, Ode Takahiro, Okada Hidehiko, Nagata Koji, Kiyohara Motosuke, Suzuki Ei-Ichiro, Kita Akiko, Wada Hitoshi, Tanokura Masaru

机构信息

Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.

Institute for Innovation, Ajinomoto Co. Inc., 1-1 Suzuki-cho, Kawasaki-ku, Kawasaki, Kanagawa 210-8681, Japan.

出版信息

Sci Rep. 2016 Feb 26;6:22127. doi: 10.1038/srep22127.

Abstract

Precise protein structure determination provides significant information on life science research, although high-quality crystals are not easily obtained. We developed a system for producing high-quality protein crystals with high throughput. Using this system, gravity-controlled crystallization are made possible by a magnetic microgravity environment. In addition, in-situ and real-time observation and time-lapse imaging of crystal growth are feasible for over 200 solution samples independently. In this paper, we also report results of crystallization experiments for two protein samples. Crystals grown in the system exhibited magnetic orientation and showed higher and more homogeneous quality compared with the control crystals. The structural analysis reveals that making use of the magnetic microgravity during the crystallization process helps us to build a well-refined protein structure model, which has no significant structural differences with a control structure. Therefore, the system contributes to improvement in efficiency of structural analysis for "difficult" proteins, such as membrane proteins and supermolecular complexes.

摘要

精确的蛋白质结构测定为生命科学研究提供了重要信息,尽管高质量的晶体并不容易获得。我们开发了一种能够高通量生产高质量蛋白质晶体的系统。利用该系统,通过磁微重力环境实现了重力控制结晶。此外,对于200多个溶液样品,可以独立地对晶体生长进行原位实时观察和延时成像。在本文中,我们还报告了两个蛋白质样品的结晶实验结果。在该系统中生长的晶体呈现出磁取向,与对照晶体相比,其质量更高且更均匀。结构分析表明,在结晶过程中利用磁微重力有助于构建精制良好的蛋白质结构模型,该模型与对照结构没有明显的结构差异。因此,该系统有助于提高对“难”蛋白质(如膜蛋白和超分子复合物)的结构分析效率。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e225/4768258/ebb51a48575f/srep22127-f1.jpg

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